The overall goal of this application is to evaluate new imaging agents for (1) real-time detection of residual cancer in the tumor bed during surgery in primary colon and lung cancers in mice to eliminate or significantly reduce the need for repeat surgeries and local recurrence, and (2) guiding resection surgeries in canine patients with naturally occurring cancer. In a pilot study, we will also test our imaging technology to improve the detection of lesions during routine colonoscopy examination. Failure to remove cancer cells during surgery is a leading risk factor for local tumor recurrence and subsequent reoperation. Rates of secondary surgeries because of positive margins range between 10-50% depending on cancer type, grade and anatomical site. For colon cancer screening, many small lesions go undetected that are only found after progressing into malignant carcinomas. Furthermore, even for larger carcinomas, colonoscopy using white light will not identify these 22% of the time. Thus, a technology that can assess whether tumor cells have been adequately removed during surgery or can identify colon lesions with higher efficiency will have a major impact on cancer therapy worldwide as more than two million people undergo cancer surgery every year. During the SBIR Phase I work, Lumicell developed several imaging agents by using our novel mathematical model to predict their performance across species. The goal was to develop high-performance, lower cost imaging agents to satisfy the needs for colon cancer screening and eventually skin cancer surgeries. These imaging agents are based upon the design of our lead candidate for sarcoma and breast cancer indications, LUM015, which has been approved by the FDA to start first-in-human clinical trials with our clinical collaborators at Duke University Medical Center. Based upon modifications to LUM015, the new imaging agents cost about 66% less to manufacture with improved yields. After testing them in our baseline mouse model for breast cancer, these new agents proved equal or better performance than LUM015. A lower cost imaging agent is mandatory for entry into several of our new target markets and will be an additional offering for the other markets. For Phase II, we propose in Aims 1 and 2 to evaluate the performance of these imaging agents in state-of-the-art genetically engineered mouse models for lung cancer and colon cancer with our collaborators at Duke University and Massachusetts General Hospital. In Aim 3, we will move forward with the selected lead candidate (based upon results from Aims 1 and 2) into a veterinary clinical trial conducted at Duke/Veterinary Specialty Hospital of the Carolinas and Tufts University Cummings School of Veterinary Medicine. Canine patients suffering from any type of cancer will be recruited to evaluate the performance of the lead candidate in naturally occurring cancers. Aim 4 is focused on reaching the endpoint of this proposal: submit an IND to support a Phase I clinical trial for colon and lung cancer surgeries and colon cancer screening using our new lead candidate.